JP4796043B2 - Periodontal disease growth inhibitor - Google Patents

Description

The present invention relates to a periodontal bacterial growth inhibitor.

  Periodontal diseases that develop due to infection with periodontal disease bacteria are roughly classified into gingivitis, periodontitis, and occlusal trauma. Recently, it is known that periodontal disease becomes chronic and causes infections such as respiratory diseases, heart diseases, diabetes, and premature birth due to periodontal disease bacteria. As an antibacterial agent against such periodontal disease bacteria, one using cocoa contained in cacao mass as an active ingredient is known (see Patent Document 1).

On the other hand, it is known that an extract of a giant grass (major tree) belonging to the genus Euphoridae exhibits an antibacterial action (see Patent Document 2).
International Publication No. 2003/99304 Pamphlet JP 2007-45754 A

  By the way, in the antibacterial agent of the said patent document 1, in order to obtain raw material cacao mass, the process of roasting cacao beans etc. are required. That is, there is a problem that even if it is obtained from a plant-derived raw material and is a periodontal disease growth inhibitor that can suppress the growth of periodontal disease bacteria, the raw material production process becomes complicated. This invention is made | formed by discovering that a weevil extract exhibits the effect | action which suppresses the growth of periodontal disease bacteria as a result of earnest research by these investigators. In addition, in the said patent document 2, the effect | action of the grasshopper extract with respect to a periodontal disease microbe is not elucidated.

An object of the present invention is to provide a periodontal bacterial growth inhibitor that can be obtained from a raw material that can avoid complication of pretreatment.

In order to achieve the above object, the periodontal disease bacterial growth inhibitor of the invention described in claim 1 contains nymphaeol-A, nymphaeol-B and nymphaeol- C as active ingredients, and the active ingredient is a lower alcohol. It is characterized by suppressing the growth of periodontal disease bacteria contained in the genus Prophyromonas by being contained as a plantain extract extracted with an extraction solvent containing .

The invention according to claim 2 is characterized in that in the periodontal disease bacteria growth inhibitor according to claim 1, the extraction solvent contains ethanol as the lower alcohol .
The invention according to claim 3 is the periodontal disease growth inhibitor of claim 1 or claim 2, wherein the periodontal disease bacteria contained in the genus Prophyromonas is Porphyromonas gingivalis. It is characterized by.
The invention according to claim 4 is the periodontal disease growth inhibitor according to any one of claims 1 to 3, wherein the periodontal disease growth inhibitor is the nymphaeol-A or the nymphaeol-B. And the concentration of the total amount of Nimpheol-C is at least 50 ppm .

ADVANTAGE OF THE INVENTION According to this invention, the periodontal disease bacteria growth inhibitor which can be obtained from the plant raw material which can avoid the complication of a pretreatment is provided.

Hereinafter, an embodiment embodying the present invention will be described in detail.
The periodontal disease bacteria growth inhibitor of this embodiment contains a plantain extract extracted with an extraction solvent containing at least an organic solvent as an active ingredient. The giant grass (big leaf tree) that is the raw material of the giant grass extract is a plant called Macaranga tanarius, and is an evergreen broad-leaved tree (six male and female) belonging to the genus Euphorbiaceae. Weow grows in Okinawa, Taiwan, southern China, the Malay Peninsula, Southeast Asia such as the Philippines, Malaysia, Indonesia, and Thailand, and northern Australia. In addition, weow grows very fast among trees and can grow in degraded areas.

  As a raw material for the plant extract, various organs and components of the plant can be used. As a raw material, a single organ or component may be used, or two or more organs or components may be mixed and used. From the viewpoint that the effect of inhibiting the growth of periodontal disease bacteria by the grasshopper extract increases, the raw material preferably contains fruits, seeds, flowers, roots, stems, stem tips, leaf blades, and secretions (wax, etc.). Since the tip of the stem includes the growth point of the stem and the leaf bud and is more flexible than the leaf blade, it is easy to perform the extraction operation efficiently. In addition, when the proportions of the respective organs with respect to the whole of the grasshopper are compared, the proportions occupied by the trunk, roots, and leaves are high. For this reason, it is industrially suitable from the viewpoint that it is easy to secure the raw material to use easy-to-handle green grass as a raw material for the extract. Such raw materials can be extracted as they are collected, crushed, crushed or ground after collection, crushed, crushed or ground after collection or drying, or crushed, crushed or ground after collection and dried. The operation can be performed. In order to perform the extraction operation efficiently, it is preferable to use a crushed raw material. For such crushing, for example, a cutter, a cutter, a crusher, or the like can be used. The shape of the crushed raw material is not particularly limited, and examples thereof include polygonal shapes such as a triangular shape and a quadrangular shape. In addition, as for the dimension of the crushed raw material, it is preferable that one side is about 1 cm. On the other hand, when preparing the pulverized raw material, for example, a mill, a crusher, a grinder or the like can be used. In preparing the ground material, a kneader, a mortar, or the like can be used.

  As an extraction solvent for extracting the wean extract from the above-mentioned raw materials, a mixed solvent of water and an organic solvent, an organic solvent such as a lower alcohol, dimethyl sulfoxide, acetonitrile, acetone, ethyl acetate, hexane, glycerin, propylene glycol and the like. Can be mentioned. Examples of the lower alcohol include methanol, ethanol, propanol, isopropanol, butanol and the like. As an organic solvent, a single species may be used, or a mixed solvent obtained by mixing a plurality of species may be used. Glycerin or propylene glycol may be blended as a solvent, a wetting agent or the like in oral hygiene products such as dentifrices and mouthwashes. Therefore, in accordance with the formulation of the grass extract using glycerin or propylene glycol as an extraction solvent, glycerin or propylene glycol can be blended as a solvent, a wetting agent or the like for oral hygiene products. Thus, it is preferable that an extraction solvent contains glycerol or propylene glycol from a viewpoint of achieving efficiency about manufacture of an oral hygiene product. When a mixed solvent of water and an organic solvent is used as the extraction solvent, the content of the organic solvent in the mixed solvent is preferably 50% by volume or more, more preferably 80% by volume or more. When the content of the organic solvent in the mixed solvent is less than 50% by volume, there is a possibility that the active ingredient contained in the grass can not be efficiently extracted. In addition, as an organic solvent, a lower alcohol is preferable and ethanol is more preferable.

In addition, you may dissolve organic salt, inorganic salt, a buffering agent, an emulsifier, dextrin, etc. in an extraction solvent.
As the extraction operation, the raw material is immersed in the extraction solvent for a predetermined time. In such an extraction operation, a stirring operation, heating, or the like may be performed as necessary to increase the extraction efficiency. Further, in order to reduce impurities extracted from the raw material, a separate water extraction operation or hot water extraction operation may be performed prior to the extraction operation. Ingredients that effectively act on the suppression of the growth of periodontal disease bacteria are nymphaeols contained in the grass. Since nymphaeols are components that are insoluble only in water, unnecessary impurities other than nymphaeols can be efficiently removed by boiling the grass with hot water, for example.

  After the extraction operation, a solid-liquid separation operation is performed to separate the grass extract and the raw material residue. As a separation method for such a solid-liquid separation operation, a known separation method such as filtration or centrifugation can be used. The obtained grass extract may be concentrated as necessary.

  In addition, by removing the extraction solvent contained in the grass extract as necessary, a solid grass extract can be obtained. Such removal of the solvent may be performed, for example, by heating under reduced pressure or by freeze-drying.

  Nymphaeols are contained in the plantain extract extracted with an extraction solvent containing at least an organic solvent. The nymphaeols include at least one selected from nymphaeol-A, nymphaeol-B, and nymphaeol-C. Nymphaeol-A is 5,7,3 ′, 4′-tetrahydroxy-6-geranylflavanone (5,7,3 ′, 4′-tetrahydroxy-6-geranylflavanone). Nymphaeol-B is 5,7,3 ′, 4′-tetrahydroxy-2′-geranylflavanone (5,7,3 ′, 4′-tetrahydroxy-2′-geranylflavanone). Nymphaeol-C is 5,7,3 ′, 4′-tetrahydroxy-6- (3 ″ ″, 3 ″ ″-dimethylallyl) -2′-geranylflavanone (5,7, 3 ′, 4′-tetrahydroxy-6- (3 ′ ″ ″, 3 ′ ″ ″-dimethylallyl) -2′-geranylflavanone).

The main component of the grasshopper extract is the above-mentioned nymphaeols, and it is speculated that these nymphaeols contribute to the inhibitory action of periodontal disease bacteria.
In addition, the plantain extract contains proporin A. Proporin A is 5,7,3 ′, 4′-tetrahydroxy-2 ′-(7 ″ -hydroxy-3 ″, 7 ″ -dimethyl-2 ″ -octenyl) -flavanone (5 7,3 ′, 4′-tetrahydroxy-2 ′-(7 ″ -hydroxy-3 ″, 7 ″ -dimethyl-2 ″ -octenyl) -flavanone). As a trace component, the moth extract contains 5,7,3 ′, 4′-tetrahydroxy-5′-geranylflavanone (isonymphaeol B: 5,7,3 ′, 4′-tetrahydroxy -5´-geranylflavanone), 5,7,3´, 4´-tetrahydroxy-5´- (7´´-hydroxy-3´´, 7´´-dimethyl-2´´-octenyl) -flavanone (5 , 7,3´, 4´-tetrahydroxy-5´- (7´´-hydroxy-3´´, 7´´-dimethyl-2´´-octenyl) -flavanone), 5,7,3´, 4´ -Tetrahydroxy-6- (7 "-hydroxy-3", 7 "-dimethyl-2" -octenyl) -flavanone (5,7,3 ", 4" -tetrahydroxy-6- (7 "" -hydroxy-3 ", 7" -dimethyl-2 "-octenyl) -flavanone), 5,7,4" -trihydroxy-3 "-(7" -hydroxy-3 ", 7" -Dimethyl-2 "-octenyl) -flavanone (5,7,4'-trihydroxy-3"-(7 ""-hydroxy-3 "", 7 "-dimetyl-2" -octenyl) -flavanone) , 5,7,4´-trihydroxy-3´-geranylflavanone (5,7,4´-trihydroxy-3´-ge ranylflavanone) and the like. Among the extracts extracted from each part of the grass, the extract extracted from the actual part (including wax) contains nymphaeol A, B, C and isonymfeol B at high concentrations. ing.

  The periodontal disease bacterial growth inhibitor may contain components other than the grass extract in a range that does not impair the growth inhibitory action. Examples of components other than the grass extract include excipients, bases, emulsifiers, stabilizers, fragrances, and sweeteners.

  The periodontal disease bacteria growth inhibitor may be liquid or solid. Although it does not specifically limit as a dosage form, For example, a powder, a powder, a granule, a tablet, a capsule, a pill, a liquid agent, an injection, etc. are mentioned.

  Periodontal diseases that develop due to infection with periodontal disease bacteria are roughly classified into gingivitis, periodontitis, and occlusal trauma. The periodontal disease growth inhibitor of this embodiment is used for treatment, prevention, etc. of periodontal disease by suppressing the growth of periodontal disease bacteria. As periodontal disease bacteria, periodontal disease bacteria contained in the genus Actinobacillus, the genus Porphyromonas, the genus Prevotella, the genus Fusobacterium and the like are known.

  Examples of periodontal disease bacteria contained in the genus Actinobacillus include Actinobacillus actinomycetemcomitans and the like.

  Periodontopathic bacteria contained in the genus Porphyromonas include Porphyromonas gingivalis, Prophyromonas asaccharolytica, Porphyromonas endontalis and the like. It is done.

  Examples of periodontal disease bacteria contained in the genus Prevotella include Prevotella intermedia, Prevotella nigrescens, Prevotella melaninogenica and the like.

  Periodontal disease bacteria contained in the genus Fusobacterium include Fusobacterium nucleatum, Fusobacterium necrophorum, Fusobacterium naviform AA (Fusbacterium navifor, etc.).

  Among such periodontal pathogens, periodontal pathogens contained in at least one of the genus Actinobacillus and the genus Porphyromonas are considered to be major causative bacteria for periodontal diseases. The periodontal disease growth inhibitor of this embodiment is excellent in the prevention and treatment of periodontal disease by suppressing the growth of periodontal disease bacteria contained in at least one of the genus Actinobacillus and Porphyromonas. Demonstrate the effect.

  Periodontal disease bacteria growth inhibitors are used as oral hygiene products, food and drink, pharmaceutical products, quasi drugs, and the like. The periodontal disease bacteria growth inhibitor is preferably used as an oral hygiene product or a food or drink from the viewpoint that the active ingredient is easily brought into contact with the periodontal tissue and the action of the active ingredient is sufficiently exhibited.

  The oral hygiene product of this embodiment contains the above-mentioned periodontal disease bacteria growth inhibitor. Examples of oral hygiene products include powder dentifrices, toothpastes, liquid dentifrices, mouth washes, gingival massage creams, topical application agents, troches, chewing gums, and dental floss. Since such an oral hygiene product can suppress the growth of periodontal disease bacteria in the oral cavity, it is effective for the prevention and treatment of periodontal disease. In such an oral hygiene product, the aerial extract is preferably contained in the range of 0.001 to 10% by mass, more preferably 0.01 to 1% by mass in terms of solid content. When the content of the lobster extract in the oral hygiene product is less than 0.001% by mass in terms of solid content, it may be difficult to enhance the action of suppressing the growth of periodontal disease bacteria. On the other hand, when the content of the grasshopper extract in the oral hygiene product exceeds 10% by mass in terms of solid content, it is difficult to further enhance the action of suppressing the growth of periodontal disease bacteria. The oral hygiene product contains an emulsifier, a solvent, a stabilizer, and the like as necessary in addition to the base material corresponding to the form.

  The food / beverage products of this embodiment are drinks or foodstuffs, and contain the said periodontal disease bacteria growth inhibitor. Examples of the beverage include carbonated drinks, tea drinks, soft drinks, alcoholic beverages, milk, coffee, fruit syrup, jelly drinks, and nutritional drinks. Examples of carbonated drinks include cider, lemon squash, and cola. Examples of tea beverages include green tea, oolong tea, and black tea. Examples of soft drinks include fruit juice drinks and mineral water. Examples of liquors include beer, sparkling liquor, sake, whiskey, shochu, and cocktails. Examples of food include yogurt, soup, curry, throat lozenge, candy, cookies, cake, Japanese confectionery, snack confectionery, syrup and the like.

  The food and drink may be health food, food for specified health use, health drink, nutritional supplement, and the like. In such foods and drinks, the grasshopper extract is preferably contained in the range of 0.001 to 10% by mass, more preferably 0.01 to 1% by mass in terms of solid content. In the case where the content of the plantain extract in the food or drink is less than 0.001% by mass in terms of solid content, it may be difficult to enhance the action of suppressing the growth of periodontal disease bacteria. On the other hand, when the content of the grasshopper extract in the food or drink exceeds 10% by mass in terms of solid content, it is difficult to further enhance the action of suppressing the growth of periodontal disease bacteria. In addition to the base material according to the form, food / beverage products contain an emulsifier, a solvent, a stabilizer, etc. as needed.

The effects exhibited by the above embodiment will be described below.
(1) The periodontal disease bacteria growth inhibitor of this embodiment contains the plantain extract extracted with the extraction solvent containing an organic solvent at least as an active ingredient. The grasshopper extract can be easily and efficiently obtained by the procedure of leafhopper extraction. And the present researchers have found that nymphaeols contained in such a plantain extract exert an action of suppressing the growth of periodontal disease bacteria. Thereby, the periodontal disease bacteria growth inhibitor of this embodiment can be obtained from the plant raw material which can avoid complication of a pretreatment.

  (2) The grasshopper grows very fast among trees and can grow in degraded areas. In this way, the grasshopper does not take time and effort for its cultivation management. In addition, the plantain extract is a plant-derived raw material, and thus has high safety. Therefore, in addition to the effect | action which suppresses the growth of a periodontal disease microbe, the periodontal disease bacteria growth inhibitor of this embodiment is excellent also about supply of a raw material, productivity, safety | security, etc.

  (3) The periodontal disease bacteria growth inhibitor of this embodiment is prevention and treatment of periodontal disease by suppressing the growth of the periodontal disease bacteria contained in at least one of the genus Actinobacillus and Porphyromonas. Demonstrates excellent effects.

  (4) Since the oral hygiene product and food / beverage products of this embodiment contain the said periodontal disease bacteria growth inhibitor, they can be obtained from the plant raw material which can avoid complication of a pretreatment. Furthermore, such oral hygiene products and foods and drinks are excellent in the supply of raw materials, productivity, safety, etc. in addition to the action of suppressing the growth of periodontal disease bacteria.

In addition, the said embodiment can also be changed and comprised as follows.
-The said periodontal disease bacteria growth inhibitor can also be comprised so that it may contain as an active ingredient at least 1 sort (s) chosen from nimfeol-A, nimpheol-B, and nimpheol-C, and may suppress the growth of a periodontal disease bacteria. That is, since the active ingredient that suppresses the growth of periodontal disease bacteria is at least one selected from nymphaeol-A, nymphaeol-B, and nymphaeol-C, these active ingredients are derived from, for example, a grasshopper extract. Okinawa propolis or the like may be derived. In addition, for example, a mixture of nimpheol derived from Okinawa propolis and nympheol derived from Euphorus extract may be used as the active ingredient.

When the active ingredient is derived from a grasshopper extract, the action and effect described in the column (2) can be obtained.
The technical idea that can be grasped from the above embodiment will be described below.

The periodontal fungus growth inhibitor according to claim 1 , wherein the plantain extract is extracted from a mixed solvent of water and an organic solvent.

-The above-mentioned periodontal pathogen growth inhibitor that suppresses the growth of at least one periodontal pathogen of Actinobacillus, Actinomycetemcomitans, and Porphyromonas gingivalis.

Next, the embodiment will be described more specifically with reference to test examples.
(Example 1)
<Preparation of Euphorus extract 1>
After thawing the fresh leaves of the collected wolfweed collected in Okinawa, the raw leaves were cut into small pieces with scissors. 30 g of the cut fresh leaves and 100 ml of the solvent were put in a tube, immersed for 2 weeks at room temperature, subjected to solvent extraction, and then filtered to obtain a grasshopper extract as a filtrate. In the solvent extraction, a mixed solvent in which ethanol and water were mixed in a volume ratio of ethanol: water = 90: 10 was used. Next, the wolf extract was freeze-dried to prepare a wolf extract, which is a solid powder contained in the wolf extract. The concentration of nymphaeol contained in this extract was calculated from the chromatogram obtained by analyzing under the following HPLC conditions, and was found to be 50% by mass. In addition, the density | concentration of this nymphaeol has shown the density | concentration which totaled nymphaeol-A, nymphaeol-B, and nymphaeol-C. In the following, the amount of nymphaeol indicates the total amount of nymphaeol-A, nymphaeol-B, and nymphaeol-C.

(HPLC conditions)
System: PDA-HPLC system (Shimadzu Corporation), LC10ADvp series, UV; SPD-10Avp, PDA; SPD-M10Avp
Column: Luna C18 (2.0 × 250mm) (Shimadzu GLC)
Solvent: A: Water (5% acetic acid), B: Acetonitrile (5% acetic acid)
Elution conditions: 0-20min
(Gradient elution; A: B = 80: 20 → A: B = 30: 70)
20-50min
(Gradient elution; A: B = 30: 70 → A: B = 0: 100)
50-60 min (A: B = 0: 100)
60-75 min (A: B = 80: 20)
Flow rate: 0.2 ml / min
PDA detection: UV 190-370nm
UV detection: UV287nm
Injection volume: 20 μl
Temperature: 40 ° C
The chromatogram of this 50% by weight sample is shown in FIG.

<Antibacterial activity test of the grasshopper extract against periodontal disease>
The following 1 and 2 strains of periodontal disease bacteria shown below were inoculated on a GAM agar medium using a platinum loop, and then cultured at 35 ° C. for 7 days.

Strain 1: Actinobacillus actinomycetemcomitans (JCM8577)
Strain 2: Porphyromonas gingivalis (JCM12257)
Thus, after growing colonies were collected from the GAM agar medium using a platinum loop, 10 ⁴ cfu / ml inoculum 1 and 2 were prepared by dissolving in physiological saline.

  A blank medium and each sample medium having final concentrations of the following values were prepared by mixing ethanol and the aerial extract obtained in Preparation 1 of the above-mentioned aerial extract in a GAM agar medium.

Blank: Ethanol 0.7%
Sample 1: Awesome extract 100ppm
Sample 2: Awesome extract 250ppm
Sample 3: Awesome extract 500ppm
A plate medium was prepared by sterilizing each medium at 115 ° C. for 15 minutes.

  After inoculating the above inoculum 1 and inoculum 2 with platinum loops on each plate, the plates were cultured at 35 ° C. for 7 days in an anaerobic system manufactured by Mitsubishi Gas Chemical Company. As a result, the concentration of psyllium extract for the plate medium in which no colony was detected is shown in Table 1 below.

表１の結果から明らかなように、上記オオバギ抽出物の調製１で得られたオオバギ抽出物は菌株１の歯周病菌の発育を２５０ｐｐｍの濃度で阻止することが確認された。また、菌株２の歯周病菌の発育に対しては、１００ｐｐｍの濃度で阻止することが確認された。なお、こうしたオオバギ抽出物の抗菌活性は、オオバギ抽出物中に含まれるニムフェオール類が有効成分として寄与することで発現されると推測される。オオバギ抽出物中のニムフェオールに換算すると、上記オオバギ抽出物は、菌株１及び菌株２をそれぞれ１２５ｐｐｍ及び５０ｐｐｍの濃度で阻止することができると推定される。こうした結果から、本試験例のオオバギ抽出物は、とくに菌株２の増殖を抑制する作用に優れることがわかる。

As is clear from the results in Table 1, it was confirmed that the plantain extract obtained in Preparation 1 of the plantain extract inhibits the growth of periodontal disease bacteria of strain 1 at a concentration of 250 ppm. Further, it was confirmed that the growth of periodontal disease bacteria of strain 2 was inhibited at a concentration of 100 ppm. In addition, it is estimated that the antibacterial activity of such a plantain extract is expressed by the contribution of nymphaeols contained in the plantain extract as an active ingredient. When converted to nympheol in the grasshopper extract, the grasshopper extract is estimated to be able to block strain 1 and strain 2 at concentrations of 125 ppm and 50 ppm, respectively. From these results, it can be seen that the grasshopper extract of this test example is particularly excellent in the action of suppressing the growth of strain 2.

(Example 2)
<Preparation 2 of the grasshopper extract>
Extract 0.2 to 0.3 g of the cut fresh leaves first in water (95 ° C.) for 30 minutes, filter this, and perform extraction in the same manner as Preparation 1 of the Oobagi extract, except that solvent extraction was performed. Went.

  First, 30 g of the cut fresh leaves were immersed in water (95 ° C.) for 30 minutes and filtered. Solvent extraction was performed by adding 100% ethanol to the remaining leaves after filtration and immersing them for 3 days. The extract was filtered to collect the lobster extract, and then the lobster extract was lyophilized to prepare a lobster extract that is a solid powder contained in the lobster extract.

The concentration of nymphaeol contained in this extract was calculated from the chromatogram obtained by analyzing under the above-mentioned HPLC conditions. As a result, it was 40% by mass.
The chromatogram of this 40% by weight sample is shown in FIG.

<Antibacterial activity test of the grasshopper extract against periodontal disease>
When the antibacterial activity test was carried out in the same manner as in Example 1, the plantain extract prepared in Example 2 also showed the same antibacterial activity.

The chromatogram which shows the result analyzed by the high performance liquid chromatography about the grasshopper extract which concerns on Example 1. FIG. The chromatogram which shows the result analyzed by the high performance liquid chromatography about the grasshopper extract which concerns on Example 2. FIG.

Claims (4)

It contains nymphaeol-A, nymphaeol-B, and nymphaeol- C as active ingredients, and the active ingredient is contained as a plantain extract extracted with an extraction solvent containing a lower alcohol, so that it belongs to the genus Prophyromonas. The periodontal disease growth inhibitor which suppresses the growth of the periodontal disease bacteria contained in . The periodontal pathogen growth inhibitor according to claim 1, wherein the extraction solvent contains ethanol as the lower alcohol . The periodontal pathogen growth inhibitor according to claim 1 or 2, wherein the periodontal pathogen contained in the genus Prophyromonas is Porphyromonas gingivalis . The said periodontal disease bacteria growth inhibitor is used so that the density | concentration of the total amount of the said nymphaeol-A, the said nymphaeol-B, and the said nymphaeol-C may be set to at least 50 ppm. The periodontal disease bacteria growth inhibitor as described in any one of these.

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